scholarly journals Evidence for the Use of Mucus Swabs to Detect Renibacterium salmoninarum in Brook Trout

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 460
Author(s):  
Tawni B. Riepe ◽  
Victoria Vincent ◽  
Vicki Milano ◽  
Eric R. Fetherman ◽  
Dana L. Winkelman
Keyword(s):  
Brook Trout ◽  
Sampling Methods ◽  
Kidney Tissue ◽  
Detection Methods ◽  
Fish Health ◽  
Wild Populations ◽  
Renibacterium Salmoninarum ◽  
Tissue Samples ◽  
Future Studies ◽  
Aquaculture Facilities

Efforts to advance fish health diagnostics have been highlighted in many studies to improve the detection of pathogens in aquaculture facilities and wild fish populations. Typically, the detection of a pathogen has required sacrificing fish; however, many hatcheries have valuable and sometimes irreplaceable broodstocks, and lethal sampling is undesirable. Therefore, the development of non-lethal detection methods is a high priority. The goal of our study was to compare non-lethal sampling methods with standardized lethal kidney tissue sampling that is used to detect Renibacterium salmoninarum infections in salmonids. We collected anal, buccal, and mucus swabs (non-lethal qPCR) and kidney tissue samples (lethal DFAT) from 72 adult brook trout (Salvelinus fontinalis) reared at the Colorado Parks and Wildlife Pitkin Brood Unit and tested each sample to assess R. salmoninarum infections. Standard kidney tissue detected R. salmoninarum 1.59 times more often than mucus swabs, compared to 10.43 and 13.16 times more often than buccal or anal swabs, respectively, indicating mucus swabs were the most effective and may be a useful non-lethal method. Our study highlights the potential of non-lethal mucus swabs to sample for R. salmoninarum and suggests future studies are needed to refine this technique for use in aquaculture facilities and wild populations of inland salmonids.

scholarly journals Ability of Real-time PCR in diagnose Differentiation Various Forms of Cutaneous Leishmaniasis: A Comparative Study with Histopathology

2019 ◽  
Author(s):  
Maryam Fekri Soofi Abadi ◽  
Meisam Fekri ◽  
alireza moradabadi ◽  
Reza Vahidi ◽  
Simin Shamsi Meymandi ◽  
...  
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Real Time ◽  
Real Time Pcr ◽  
Relative Number ◽  
Parasite Load ◽  
Detection Methods ◽  
Tissue Samples ◽  
Microscopic Evaluation ◽  
Histopathological Evaluation ◽  
Histopathological Studies

Abstract objective: Histopathological studies suggest that parasite load is different between acute and chronic forms of cutaneous leishmaniasis (CL). However, highly sensitive detection methods are still needed to distinguish different forms of leishmaniasis. In the present study, we developed a quantitative real-time polymerase chain reaction (PCR) to detect and quantify leishmania tropica parasites in paraffin-embedded tissue samples. Results: The ability of real-time PCR for leishmania detection was higher than histopathological evaluation. The parasite loads were quantified by qPCR assay and microscopic evaluation were highly correlated ( r =0.598; P <0.001). Among patients, the parasite load was inversely correlated with disease duration (acute CL lesions had very higher parasite loads than chronic CL lesions), but there was no difference in parasite load according to the patients’ age and sex as well as location of the lesions. In contrast to Ridley scoring system (P<0.001), there were no statistically significant differences in the relative number of parasites among the lupoid and non-lupoid forms of chronic lesions in real-time PCR (P=0.549), which indicates the superiority of histopathological evaluation in CL forms differentiation.

scholarly journals The Effect of Zinc and Melatonin Supplementation on Lipid Peroxidation of Various Tissues of Rats with by DMBA-Induced Breast Cancer

2020 ◽  
Vol 11 (1) ◽  
pp. 7580-7588
Keyword(s):  
Breast Cancer ◽  
Lipid Peroxidation ◽  
Kidney Tissue ◽  
Female Rats ◽  
Control Group ◽  
Tissue Samples ◽  
Cancer Group ◽  
Group 5 ◽  
Group 2 ◽  
Group Control Group

The purpose of this study was to investigate the effects of zinc and melatonin administration on lipid peroxidation in various tissues in DMBA-induced breast cancer in female rats. A total of 42 recently weaned Wistar rats were divided into 5 groups in the study: Group 1 (Control), Group 2 (DMBA Control), Group 3 (DMBA+Zinc), Group 4 (DMBA+Melatonin), Group 5 (DMBA+Melatonin, and Zinc). MDA (malondialdehyde) and GSH (glutathione) levels were determined via the spectrophotometric method in the lung, liver, spleen, pancreas, and kidney tissue samples taken from experimental animals. The highest lung, liver, spleen, pancreas, and kidney tissue MDA levels were obtained in the DMBA-induced breast cancer group control group (G2) (p<0.05). MDA levels in DMBA+Zinc (G3), DMBA+Melatonin (G4), and DMBA+Melatonin and Zinc (G5) were significantly lower than group 2 (p<0.05). Similarly, lung, liver, spleen, pancreas, and kidney tissue GSH levels of DMBA+Zinc (G3), DMBA+Melatonin (G4), and DMBA+Melatonin and Zinc (G5) were significantly higher than that of Group 2 (p<0.05). The findings of the study indicated that increased lung, liver, spleen, pancreas, and kidney damage in DMBA-induced breast cancer is suppressed with the supplementation of zinc, melatonin, and combined zinc and melatonin.

Carvacrol attenuates amikacin-induced nephrotoxicity in the rat

2021 ◽  
Author(s):  
Atta Mohammad Dost ◽  
Mehmet Gunata ◽  
Onural Ozhan ◽  
Azibe Yildiz ◽  
Nigar Vardi ◽  
...  
Keyword(s):  
Inflammatory Cell ◽  
Kidney Tissue ◽  
Control Group ◽  
Histopathological Changes ◽  
Sprague Dawley ◽  
Tissue Samples ◽  
Sprague Dawley Rats ◽  
Before And After ◽  
Per Oral

Abstract Amikacin (AK) is frequently used in the treatment of gram-negative and some gram-positive infections. However, its use is limited due to nephrotoxicity due to the increase in reactive oxygen radicals. The aim of this study was to investigate the role of carvacrol (CAR) against AK-induced nephrotoxicity in rats. Thirty-two Sprague Dawley rats were randomly divided into four groups as control (Vehicle), AK (400 mg/kg), CAR + AK (80 mg/kg CAR + 400 mg/kg AK), and AK + CAR (400 mg/kg AK + 80 mg/kg CAR) groups. AK and CAR were administered via intramuscular and per-oral for 7 days, respectively. Blood and kidney tissue samples were taken at the end of the experiment. Renal function and histopathological changes were compared, and the relevant parameters of oxidative stress and inflammation were detected. Histopathological findings (necrotic changes and dilatation and inflammatory cell infiltration) significantly increased in the AK group compared to the control group. Also, the rats in the AK group lost weight significantly. It was found that CAR treatment before and after AK significantly improved nephrotoxicity histopathologically (p < 0.05). However, this improvement was not detected biochemically. These results show that CAR treatment before and after AK improves nephrotoxicity in the histopathological level.

The Effect of Thymol on Renal Toxicity Induced by Mercury Chloride in Rats

2021 ◽  
Author(s):  
Hamid Reza Jamshidi ◽  
Faezeh Taheri
Keyword(s):  
Mercuric Chloride ◽  
Renal Toxicity ◽  
Corn Oil ◽  
Kidney Tissue ◽  
Control Group ◽  
Mercury Chloride ◽  
Protective Effects ◽  
Tissue Samples ◽  
Significant Difference ◽  
Anti Oxidant

Background and Aims: Mercuric chloride is highly toxic once absorbed into the bloodstream, especially the kidneys in which it is collected. Mercury chloride increases hydrogen peroxide and enhances the destruction of protective enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPX), leading to oxidative stress. Besides, thymol has anti-oxidant effects and can increase the activity of SOD and GPX. This study aims to evaluate the efficacy of thymol on mercury chloride-induced toxicity. Materials and Methods: In this study, 30 rats, consisting of 6 groups of 5, were used. Control group receiving a single dose of 0.5 mg/kg mercuric chloride for 15 days, third, fourth, and fifth group received intraperitoneal injection of mercuric chloride at a dose of 0.5 mg/kg for 15 days plus thymol at a dose of 10, 30, 50 mg/kg. The sixth group received mercuric chloride at a dose of 0.5 mg/kg for 15 days plus thymol at 30 mg/kg per day for ten days. Results: Results showed a significant difference in the activity of catalase enzyme in kidney tissue samples test. According to the results of SOD, there is a significant difference between the group of corn oil and the group of mercury chloride and between the group of mercury chloride and the group that receives thymol at a dose of 10, 30, 50 mg/kg (p ≤ 0.05). Conclusions: It can be concluded that mercury chloride-induced kidney toxicity and thymol have anti-oxidant protective effects for SOD and GPX.

scholarly journals P0022INNATE IMMUNE RESPONSES ELICITED BY TOLL-LIKE RECEPTORS ARE POSSIBLY INVOLVED IN THE PATHOGENESIS OF NEPHROTIC SYNDROME

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Eriko Tanaka ◽  
Ichiro Hada ◽  
Naoaki Mikami ◽  
Kunimasa Yan
Keyword(s):  
Innate Immunity ◽  
Nephrotic Syndrome ◽  
Immune System ◽  
Innate Immune System ◽  
Expression Profiles ◽  
Kidney Tissue ◽  
Innate Immune ◽  
Rna Expression ◽  
Toll Like Receptors ◽  
Tissue Samples

Abstract Background and Aims Pathogenesis of idiopathic nephrotic syndrome (INS) is yet to be fully elucidated. Immunological disorders are reported to be involved in the etiology of INS. Due to the efficacy of immunosuppressant agents such as calcineurin inhibitor and rituximab in treating nephrotic syndrome, aberrant activation of the acquired immune system through T and B cells are considered to be the underlying pathogenic mechanisms of INS. Nevertheless, there is a possibility that the innate immune system plays a key role in INS pathogenesis. This study aims to investigate the involvement of innate immunity in INS pathogenesis by examining the expressions of toll-like receptors (TLRs). Method Kidney tissue samples from two INS patients were collected at two points of time: the first biopsy was performed during nephrosis and the second during remission. Total RNA was extracted from the kidney tissue samples, and RNA-sequencing was performed to investigate RNA expression profiles. The differences between RNA expression profiles of TLRs and molecules related to TLR pathways in the tissue samples collected during nephrosis and remission were analyzed. Results There was a significant decrease in RNA expression of TLR9 and TLR10 during remission compared to nephrosis: fold change in each patient was -2.12 and -2.12 for TLR9, and -2.51 and -2.09 for TLR10. RNA expression of TLR8 also decreased: fold change in each patient was -1.19 and -1.75. There were no significant changes in the RNA expression profiles of TLR1, 2, 3, 4, 5, 6, and 7. In addition, there were no differences in the RNA expression profiles of MYD88, IRAK family, and TRAF family molecules that are associated with TLR pathways. However, RNA expressions of IL6, IL1B, IL12B, and TNF, as well as the cytokines controlled by TLR8 and TLR9 pathways, which were activated during nephrosis, disappeared or decreased during remission. Conclusion The involvement of the innate immune system in the pathogenesis of nephrotic syndrome has been suggested in some reports. Based on the fact that the onset or recurrence of nephrosis is triggered by non-specific viral infection, it is highly possible that innate immunity is involved in the pathogenesis of nephrotic syndrome. TLRs play a key role in innate immunity as they elicit the innate immune system after detecting pathogens, induce inflammatory cytokine production, and trigger signaling pathways that activate lymphocytes via maturation of dendritic cells. Specifically, TLR8, 9, and 10 mediate pathways of the first immune response to viral infections. Our study reveals that TLRs play a pivotal role in innate immunity associated with renal tissue during the onset of nephrosis.

Evaluation of Environmental Sampling Methods and Rapid Detection Assays for Recovery and Identification of Listeria spp. from Meat Processing Facilities

2009 ◽  
Vol 72 (4) ◽  
pp. 696-701 ◽  
Author(s):  
JOVANA KOVAČEVIĆ ◽  
VALERIE M. BOHAYCHUK ◽  
PABLO ROMERO BARRIOS ◽  
GARY E. GENSLER ◽  
DEANA L. ROLHEISER ◽  
...  
Keyword(s):  
Sampling Methods ◽  
Culture Method ◽  
Detection Methods ◽  
Lower Sensitivity ◽  
Meat Processing ◽  
Cotton Swab ◽  
Conventional Culture ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
Labor Inputs

Studies that isolated Listeria spp. from the environment of two meat processing facilities were conducted. Samples were collected in the processing environment of the facilities with three different sampling methods (cotton swab, sterile sponge, and composite-ply tissues) to evaluate their ability to recover Listeria spp. A total of 240 samples for each sampling method were collected and tested. The cotton swab method of sampling was significantly (P &lt; 0.01) less efficient in recovery of Listeria spp. than the sterile-sponge and composite-ply tissue methods, which were similar (P &gt; 0.05) in their ability to recover Listeria spp. The specificity and sensitivity of four detection methods (conventional culture, Petrifilm Environmental Listeria Plates [ELP], lateral-flow immunoprecipitation [LFI], and automated PCR) were evaluated for identification of Listeria spp. Facilities were visited until a minimum of 100 positive and 100 negative samples per detection method were collected. The LFI and PCR methods were highly sensitive (95.5 and 99.1%, respectively) and specific (100%) relative to the culture method. The ELP method was significantly less efficient (P &lt; 0.01) than LFI and PCR in detection of Listeria spp., with lower sensitivity (50.6%) and specificity (91.5%). Kappa values indicated excellent agreement of the LFI and PCR assays and moderate agreement of the ELP method to the culture method. Overall, ELP was easy to use but less efficient in detection of Listeria spp. from environmental samples, while the LFI and PCR methods were found to be excellent alternatives to culture, considering performance and time and labor inputs.

Protein profiling of microdomains purified from renal cell carcinoma and normal kidney tissue samples

2012 ◽  
Vol 8 (4) ◽  
pp. 1007-1016 ◽  
Author(s):  
F. Raimondo ◽  
L. Morosi ◽  
C. Chinello ◽  
R. Perego ◽  
C. Bianchi ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Kidney Tissue ◽  
Protein Profiling ◽  
Normal Kidney ◽  
Tissue Samples ◽  
Normal Kidney Tissue

scholarly journals Comparison of detection methods for Salmonella enterica shedding among reptilian patients at a veterinary teaching hospital

2019 ◽  
Vol 32 (1) ◽  
pp. 118-123
Author(s):  
Anna C. Fagre ◽  
Kristy L. Pabilonia ◽  
Matthew S. Johnston ◽  
Paul S. Morley ◽  
Brandy A. Burgess
Keyword(s):  
Salmonella Enterica ◽  
Sampling Methods ◽  
The United States ◽  
Cross Sectional Study ◽  
Detection Methods ◽  
Sample Type ◽  
Cross Sectional ◽  
Significant Disagreement ◽  
Culture Positive ◽  
Positive Results

In the United States, ~1.4 million sporadic human Salmonella enterica infections occur annually, with an estimated 6% attributable to reptile exposure. Detection of Salmonella in reptiles can be challenging given the limitations among detection methods. We evaluated sampling and detection methods for S. enterica in a cross-sectional study of reptilian patients ( n = 45) over the course of 13 mo. Two sampling methods (cloacal swabs, electrostatic cloth body-feet samples) and 3 detection methods (enriched culture, lateral flow immunoassay [LFI], real-time PCR) were compared using McNemar and Fisher exact tests. Results varied by species, sample type, and detection method. In total, 14 of 45 (33%) patients were positive by culture, 10 of 45 (22%), and/or 13 of 45 (29%) by rtPCR. Among rtPCR-positive results, cloacal swabs (12 of 45 [27%]) resulted in a higher detection than body-feet wipes (4 of 45 [9%]; p = 0.01). Among culture-positive results, shedding was most commonly detected after additional incubation at room temperature when testing cloacal swabs (9 of 45 [20%]). However, there was significant disagreement between sampling methods (cloacal vs. body-feet; p = 0.03). No samples were positive by LFI. In general, cloacal swabs yielded the highest test-positive rates, irrespective of testing method. Our study highlights the importance of using detection methods optimized for the sample being tested.

Sampling Methods and Frozen Storage of Samples for Detection of Campylobacter jejuni on Freshly Processed Broiler Carcasses

1983 ◽  
Vol 46 (6) ◽  
pp. 510-513 ◽  
Author(s):  
L. C. BLANKENSHIP ◽  
S. E. CRAVEN ◽  
J. Y. CHIU ◽  
G. W. KRUMM
Keyword(s):  
Campylobacter Jejuni ◽  
Sampling Method ◽  
Sampling Methods ◽  
Frozen Storage ◽  
Test Tube ◽  
Tissue Samples ◽  
Defibrinated Sheep Blood ◽  
Cryoprotective Agents ◽  
Plastic Bags ◽  
Sampling Locations

Swab, rinse and excision sampling methods are commonly used for detection of microorganisms on poultry carcasses. Swabbing has been the most frequently reported sampling method for Campylobacter jejuni on poultry. We evaluated the three methods for C. jejuni detection on freshly processed poultry in the following ways: (a) the interior and exterior surfaces of half of a carcass were each thoroughly rubbed with separate swabs which were combined in a test tube containing 2 ml of appropriate medium; (b) 25 g of skin and tissue samples from neck and abdominal opening cut areas were deposited in a stomacher bag with 5 ml of brucella broth (BB) and stomached for 2 min; and (c) half carcasses were shaken for 1 min with 100 ml BB in plastic bags. One drop of each sample was streaked for isolation on brucella agar containing 10% defibrinated sheep blood and Skirrow antibiotics. Isolates were identified by microscopy and appropriate cultural tests. All three sampling techniques were essentially equivalent for detection of C. jejuni on fresh carcasses. However, when samples were stored frozen for 7 to 10 d to simulate transport conditions from sampling locations to the laboratory, the incidence of detection was significantly reduced. Use of cryoprotective agents was an effective method to preserve swab samples during frozen storage.

Exploring the advantages and limitations of sampling methods commonly used in research facilities for zebrafish health inspections

2019 ◽  
Vol 54 (4) ◽  
pp. 373-385 ◽  
Author(s):  
David Marancik ◽  
Jason Collins ◽  
Josephine Afema ◽  
Christian Lawrence
Keyword(s):  
Sample Size ◽  
Pathogen Detection ◽  
Sampling Methods ◽  
Diagnostic Testing ◽  
Scientific Integrity ◽  
Fish Health ◽  
Fish Welfare ◽  
Prevalence Level ◽  
Pathogen Prevalence ◽  
Research Facilities

Examining zebrafish populations for the presence of disease is an integral component of managing fish health in research facilities. Currently, many different strategies are used for zebrafish fish health inspections, which is a scenario that may result in subjective and biased diagnostic evaluations. The goal of this study was to compare the success of pathogen detection between a sample size of randomly selected fish ( n = 60) that provides 95% confidence in pathogen detection based on a presumed pathogen prevalence level ≥5%, and other subpopulations and sample numbers commonly submitted for diagnostic testing within a 1000 tank, 30,000 fish, recirculating research system. This included fish collected from a sump tank ( n = 53), sentinel fish ( n = 11), and fish that were found moribund or freshly dead ( n = 18). Additionally, five fish from each subpopulation were collected for histopathologic examination. A second study used retrospective data to examine pathogen distribution between systems ( n = 2−5) in multi-system facilities ( n = 5) using a sample size of 60 fish per system. For the pathogens detected, results supported the use of representative sample numbers rather than smaller numbers of populations considered more at risk. The exception to this is for the moribund/mortality group, which may be a resource for targeted surveillance of select pathogens. Each system within multi-system facilities should be considered separate units in terms of fish health inspections and biosecurity. Development of these evidence-based standards for fish health inspections in zebrafish systems enhances fish welfare, provides identification of potentially zoonotic pathogens, and ensures scientific integrity and reproducibility of research results.

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